Comparison of the Donor Age-Dependent and In Vitro Culture-Dependent Mesenchymal Stem Cell Aging in Rat Model

Clinical experiments suggest that mesenchymal stem cells (MSCs) may be useful for tissue repair therapies or treatment of the autoimmune disorders. There is still lack of consensus concerning the age limit of MSC donors, majority of researchers suggest the autologous MSC therapies of patients not exceeding age limit of 55-60 yrs. The purpose of our study was to compare the selected parameters of MSCs from adipose tissue (adipose stem cell, ASC) collected from young and old rats of ages corresponding to patient’s ages 25 yrs. and 80 yrs., respectively. The differences of parameters of ASCs from young and old animals were compared with the differences between ASCs from short-term (3 passage) and long-term (30 passage) in vitro culture. Cell morphology, surface marker expression, growth potential, metabolic activity, β-galactosidase activity, clonogenic potential, angiogenic potential, and differentiation ability of ASCs from young and aged animals and from in vitro cultures at 3rd and 30th passages were compared and analyzed. It may be concluded that ASCs may be applied for autologous transplantations in aged patients. Comparison of ASC aging dynamics depending on host aging or in vitro culture duration suggests that long-term in vitro culture may affect ASCs more than natural aging process of their host. We suggest that ASCs expanded in vitro prior to their clinical use must be carefully screened for the possible aging effects resulting not only from donor age, but from the duration of their in vitro culture.

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849 A Long-Term, Mesenchyme-Free In Vitro Culture System of Colonic Epithelial Cells Retaining Stem Cell Compartment With Proliferation/Differentiation Properties

Gastroenterology ◽

10.1016/s0016-5085(10)60532-x ◽

2010 ◽

Vol 138 (5) ◽

pp. S-116

Author(s):

Shiro Yui ◽

Tetsuya Nakamura ◽

Tomohiro Mizutani ◽

Yasuhiro Nemoto ◽

Ryuichi Okamoto ◽

...

Keyword(s):

Stem Cell ◽

Epithelial Cells ◽

In Vitro Culture ◽

Culture System ◽

Cell Compartment ◽

Colonic Epithelial Cells ◽

Stem Cell Compartment ◽

In Vitro Culture System

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Expression Profile of New Marker Genes Involved in Differentiation of Canine Adipose-Derived Stem Cells into Osteoblasts

International Journal of Molecular Sciences ◽

10.3390/ijms22136663 ◽

2021 ◽

Vol 22 (13) ◽

pp. 6663

Author(s):

Maurycy Jankowski ◽

Mariusz Kaczmarek ◽

Grzegorz Wąsiatycz ◽

Claudia Dompe ◽

Paul Mozdziak ◽

...

Keyword(s):

Stem Cells ◽

In Vitro Culture ◽

Osteogenic Differentiation ◽

Molecular Mechanisms ◽

Marker Genes ◽

P Value ◽

Illumina Platform

Next-generation sequencing (RNAseq) analysis of gene expression changes during the long-term in vitro culture and osteogenic differentiation of ASCs remains to be important, as the analysis provides important clues toward employing stem cells as a therapeutic intervention. In this study, the cells were isolated from adipose tissue obtained during routine surgical procedures and subjected to 14-day in vitro culture and differentiation. The mRNA transcript levels were evaluated using the Illumina platform, resulting in the detection of 19,856 gene transcripts. The most differentially expressed genes (fold change >|2|, adjusted p value < 0.05), between day 1, day 14 and differentiated cell cultures were extracted and subjected to bioinformatical analysis based on the R programming language. The results of this study provide molecular insight into the processes that occur during long-term in vitro culture and osteogenic differentiation of ASCs, allowing the re-evaluation of the roles of some genes in MSC progression towards a range of lineages. The results improve the knowledge of the molecular mechanisms associated with long-term in vitro culture and differentiation of ASCs, as well as providing a point of reference for potential in vivo and clinical studies regarding these cells’ application in regenerative medicine.

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“Empowering” Cardiac Cells via Stem Cell Derived Mitochondrial Transplantation- Does Age Matter?

International Journal of Molecular Sciences ◽

10.3390/ijms22041824 ◽

2021 ◽

Vol 22 (4) ◽

pp. 1824

Author(s):

Matthias Mietsch ◽

Rabea Hinkel

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Treatment Strategies ◽

Cardiac Cells ◽

Aging Effects ◽

Beneficial Effects ◽

Micro Rnas ◽

New Treatment

With cardiovascular diseases affecting millions of patients, new treatment strategies are urgently needed. The use of stem cell based approaches has been investigated during the last decades and promising effects have been achieved. However, the beneficial effect of stem cells has been found to being partly due to paracrine functions by alterations of their microenvironment and so an interesting field of research, the “stem- less” approaches has emerged over the last years using or altering the microenvironment, for example, via deletion of senescent cells, application of micro RNAs or by modifying the cellular energy metabolism via targeting mitochondria. Using autologous muscle-derived mitochondria for transplantations into the affected tissues has resulted in promising reports of improvements of cardiac functions in vitro and in vivo. However, since the targeted treatment group represents mainly elderly or otherwise sick patients, it is unclear whether and to what extent autologous mitochondria would exert their beneficial effects in these cases. Stem cells might represent better sources for mitochondria and could enhance the effect of mitochondrial transplantations. Therefore in this review we aim to provide an overview on aging effects of stem cells and mitochondria which might be important for mitochondrial transplantation and to give an overview on the current state in this field together with considerations worthwhile for further investigations.

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Effect of Small Molecule Supplements during In Vitro Culture of Mouse Zygotes and Parthenogenetic Embryos on Hypoblast Formation and Stem Cell Derivation

Stem Cell Reviews and Reports ◽

10.1007/s12015-012-9382-7 ◽

2012 ◽

Vol 8 (4) ◽

pp. 1088-1097

Author(s):

K. Versieren ◽

M. Van der Jeught ◽

T. O’Leary ◽

G. Duggal ◽

J. Gerris ◽

...

Keyword(s):

Stem Cell ◽

In Vitro Culture ◽

Small Molecule ◽

Mouse Zygotes ◽

Stem Cell Derivation ◽

Parthenogenetic Embryos

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Slush nitrogen vitrification of human ovarian tissue does not alter gene expression and improves follicle health and progression in long-term in vitro culture

Fertility and Sterility ◽

10.1016/j.fertnstert.2018.08.020 ◽

2018 ◽

Vol 110 (7) ◽

pp. 1356-1366 ◽

Cited By ~ 3

Author(s):

Vincenza Barbato ◽

Roberto Gualtieri ◽

Teresa Capriglione ◽

Maria Michela Pallotta ◽

Sabrina Braun ◽

...

Keyword(s):

Gene Expression ◽

In Vitro Culture ◽

Ovarian Tissue ◽

Human Ovarian Tissue

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An in vitro cellular system modelling progressive human adipose-derived stem cell aging

Science Bulletin ◽

10.1016/j.scib.2018.01.005 ◽

2018 ◽

Vol 63 (5) ◽

pp. 272-274 ◽

Cited By ~ 1

Author(s):

Yanghua Shi ◽

Lian Wang ◽

Yichang Li ◽

Congdi Xu ◽

Xiaowen Shao ◽

...

Keyword(s):

Stem Cell ◽

Cellular System ◽

Cell Aging ◽

System Modelling ◽

Stem Cell Aging ◽

Adipose Derived Stem Cell

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Single clonal glandular stem cells derived from human parotid glands do not attain malignant phenotype during long-term in vitro culture

Neoplasma ◽

10.4149/neo_2021_210302n272 ◽

2021 ◽

Author(s):

Jung-Hwa Moon ◽

Hye-Ryun Kim ◽

Jae-Yol Lim ◽

Young-Chang Lim

Keyword(s):

Stem Cells ◽

In Vitro Culture ◽

Parotid Glands ◽

Malignant Phenotype

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Long-Term Ex Vivo Maintenance and Expansion of Transplantable Human Hematopoietic Stem Cells

Blood ◽

10.1182/blood.v94.5.1623 ◽

1999 ◽

Vol 94 (5) ◽

pp. 1623-1636 ◽

Cited By ~ 72

Author(s):

Chu-Chih Shih ◽

Mickey C.-T. Hu ◽

Jun Hu ◽

Jeffrey Medeiros ◽

Stephen J. Forman

Keyword(s):

Stem Cells ◽

Hematopoietic Stem Cells ◽

In Vitro Culture ◽

Culture System ◽

Ex Vivo ◽

Human Stem Cells ◽

Hematopoietic Stem ◽

In Vitro Culture System

Abstract We have developed a stromal-based in vitro culture system that facilitates ex vivo expansion of transplantable CD34+thy-1+ cells using long-term hematopoietic reconstitution in severe combined immunodeficient-human (SCID-hu) mice as an in vivo assay for transplantable human hematopoietic stem cells (HSCs). The addition of leukemia inhibitory factor (LIF) to purified CD34+ thy-1+ cells on AC6.21 stroma, a murine bone marrow–derived stromal cell line, caused expansion of cells with CD34+ thy-1+ phenotype. Addition of other cytokines, including interleukin-3 (IL-3), IL-6, granulocyte-macrophage colony-stimulating factor, and stem cell factor, to LIF in the cultures caused a 150-fold expansion of cells retaining the CD34+ thy-1+ phenotype. The ex vivo–expanded CD34+ thy-1+ cells gave rise to multilineage differentiation, including myeloid, T, and B cells, when transplanted into SCID-hu mice. Both murine LIF (cannot bind to human LIF receptor) and human LIF caused expansion of human CD34+ thy-1+ cells in vitro, suggesting action through the murine stroma. Furthermore, another human HSC candidate, CD34+ CD38− cells, shows a similar pattern of proliferative response. This suggests thatex vivo expansion of transplantable human stem cells under this in vitro culture system is a general phenomenon and not just specific for CD34+ thy-1+ cells.

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